Korean Journal of Veterinary Service (한국동물위생학회지)

The Korean Society of Veterinary Service (한국동물위생학회)
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A triplex real-time PCR assay for simultaneous and differential detection of Bordetella bronchiseptica, Mycoplasma cynos, and Mycoplasma canis in respiratory diseased dogs

  • Gyu-Tae Jeon (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University ) ;
  • Jong-Min Kim (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University ) ;
  • Jeong-Hyun Park (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University ) ;
  • Hye-Ryung Kim (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University ) ;
  • Ji-Su Baek (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University ) ;
  • Hyo-Ji Lee (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University ) ;
  • Yeun-Kyung Shin (Foreign Animal Disease Division, Animal and Plant Quarantine Agency) ;
  • Oh-Kyu Kwon (Foreign Animal Disease Division, Animal and Plant Quarantine Agency) ;
  • Hae-Eun Kang (Foreign Animal Disease Division, Animal and Plant Quarantine Agency) ;
  • Soong-Koo Kim (Gyeongsangbuk-do Veterinary Service Laboratory) ;
  • Jung-Hwa Kim (Gyeongsangbuk-do Veterinary Service Laboratory) ;
  • Young-Hwan Kim (Gyeongsangbuk-do Veterinary Service Laboratory) ;
  • Choi-Kyu Park (College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University )
  • Received : 2023.02.17
  • Accepted : 2023.03.20
  • Published : 2023.03.30
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Abstract

Bordetella (B.) bronchiseptica, Mycoplasma (M.) cynos, and M. canis are the major bacterial pathogens that cause canine infectious respiratory disease complex (CIRDC). In this study, we developed a triplex real-time polymerase chain reaction (tqPCR) assay for the differential detection of these bacteria in a single reaction. The assay specifically amplified three bacterial genes with a detection limit of below 10 copies/reaction. The assay showed high repeatability and reproducibility, with coefficients of intra- and inter-assay variations of less than 1%. The diagnostic results of the assay using 94 clinical samples from household dogs with CIRDC clinical signs, the prevalence of B. bronchiseptica, M. cynos, and M. canis was 22.3%, 18.1%, and 20.2%, respectively, indicating that the diagnostic sensitivity was comparable to those of previously reported qPCR assays. The dual infection rate of B. bronchiseptica and M. cynos, B. bronchiseptica and M. canis, and M. cynos and M. canis was 5.3%, 7.4%, and 3.1%, respectively. Moreover, the triple infection rate of B. bronchiseptica, M. cynos, and M. canis was 2.1%. These results indicate that coinfections with B. bronchiseptica, M. cynos, and M. canis have frequently occurred in the Korean dog population. The newly developed tqPCR assay in the present study will be a useful tool for etiological and epidemiological studies on these three CIRDC-associated bacterial pathogens. The prevalence and coinfection data revealed through this study will contribute to expanding knowledge on the epidemiology of CIRDC in the recent Korean dog population.

Keywords

Acknowledgement

This work was supported by the fund (Z-1543085-2022-23-03) by the Research of Animal and Plant Quarantine Agency, Republic of Korea.

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